【作者】 张宁；

【导师】 曹光群；

【作者基本信息】 江南大学 ， 化学工艺， 2008， 硕士

【摘要】 本课题主要对金银花叶中含有的抑菌活性物质进行了研究,包括金银花叶有效成分的提取,提取工艺的优化,抗菌谱及最低抑菌浓度的测定,探讨了抑菌活性物质的稳定性并进一步对抑菌活性物质进行分离纯化,最后对金银花叶提取物作了一些应用研究。对香樟叶、红花、金银花和金银花叶四种常见植物分别通过水和乙醇水溶液回流提取、浓缩、低温烘干。采用抑菌圈法测定各提取物的抑菌活性。分别测定它们的水提物和醇提物的抑菌活性,得到金银花叶提取物的抑菌效果强于其它几种植物提取物。采用单因素和正交实验法,考察了乙醇浓度、液料比、提取时间和提取温度四个因素对抑菌试验效果的影响,得到最佳实验条件是:金银花叶黄酮类化合物水提物的最佳提取工艺条件是液料比(V/W)14:1、提取温度70℃、提取时间4h;醇提物的最佳提取工艺条件是乙醇浓度60%,液料比(V/W)12:1,提取温度60℃,提取时间4h。将金银花叶用水和乙醇水溶液分别提取后,用抑菌圈法进行抑菌实验,得到抗菌谱;采用线性回归法快速准确地得出乙醇提取物对金黄色葡萄球菌和大肠杆菌的最低抑菌浓度。金银花叶乙醇提取物对金黄色葡萄球菌的最低抑菌浓度(MIC)为0.88%;对大肠杆菌的最低抑菌浓度(MIC)为1.21%。对金银花叶提取物抑菌活性做了稳定性测试,结果表明金银花叶提取物的抑菌活性具有较好的热稳定性;对紫外光照射稳定;对菌体的抑制能力随pH值的升高而有所下降,其中酸性条件下抑菌活性较强,在pH6-7之间,缓冲液对提取物的抑菌活性没有影响,并且提取物的抑菌效果好。对活性物质进一步分析,首先将金银花叶提取物分为石油醚部、氯仿部、乙酸乙酯部、正丁醇部和水部等不同极性溶剂萃取物,经抑菌活性比较,确定乙酸乙酯部的抑菌活性最强。乙酸乙酯部经薄层层析,展开后有多个斑点,用1%的AlCl3显色,紫外灯光下显黄绿色,推测抑菌组分属于黄酮类化合物。乙酸乙酯部分上聚酰胺和硅胶柱分离,得到抑菌效果最好的组分,通过HPLC和LC-MS分析得知带糖苷的木犀草素是金银花叶主要抑菌有效成分之一。把金银花叶提取物加入到化妆品中作测试,结果表明金银花叶提取物具有很好的防腐和抗氧化效果,可以在化妆品中广泛应用。更多还原

【Abstract】 In this paper, extract of Honeysuckle’s Leaves was tested for its antimicrobial activities, including investigation of its antimicrobial spectrum, MIC value. The processing conditions of leaching active substances from Honeysuckle’s Leaves were optimized with the orthogonal design test. The sensitivity to universal factors of the extract was studied. At the same time, the isolation and purification of the antimicrobial components of Honeysuckle’s Leaves were also studied .The water and ethanol extracts of Camphotree Leaves, safflower, Honeysuckle and its leaves were obtained by refluxing with both water and ethanol, concentration and drying, respectively. The antimicrobial activities of water and ethanol extracts against bacteria were investigated. The results showed that extract of Honeysuckle’s leaves has the stronger inhibition than others. The optimum extraction conditions of leaching active substances determined by the single factor experiment and the orthogonal design test method were as follows: The optimum extraction conditions of ethanol extract was obtained as the concentration of ethanol was 60%; temperature for extracting was 60℃with 4h, the ratio of material and solvent was l2:1. The optimum extraction conditions of water extract was obtained as the temperature for extracting was 70℃with 4h, the ratio of material and solvent was 14:1.The stability to temperature, acid, alkali and ultraviolet of the extract of Honeysuckle’s leaves were also studied. The result showed the extract was very stable to temperature and ultraviolet. And the antibacterial activities were better when pH< 7.All extracts obtained in different fractions by using different polarities of solvents such as petroleum ether, chloroform, ethyl acetate, normal butyl alcohol and water were compared through the testing of their antibacterial activities. The results showed the antibacterial activities of ethyl acetate were strongest. To track antimicrobial components in the extracts of ethyl acetate in which many different antibacterial points appeared after diffusion. The qualitative analysis of extracting sample is conducted by the specific reaction of total flavonoids with AlCl3 and so on, It is confirmed that the extraction material is flavonoids.The extracts of ethyl acetate of Honeysuckle’s leaves by polyamide columnchromatography and silica gel columnchromatography have good antibacterial activity. Antibacterial component was determined by HPLC and LC-MS. The results showed that one of antibacterial components was luteolin with a galactoside.The extract of Honeysuckle’s leaves was applied in cosmetics. All the results confirmed that extract of Honeysuckle’s leaves has the antimicrobial effect and the activity of antioxidation. So it can be widely used in cosmetics.更多还原